Injectable ready-to-use solutions containing an antitumor anthracycline glycoside

ABSTRACT

A sealed glass container containing therein a stable, injectable, sterile, pyrogen-free doxorubicin anti-tumor composition in a solution which consists essentially of a physiologically acceptable salt of doxorubicin dissolved in a physiologically acceptable solvent therefor, wherein said solution has not been reconstituted from a lyophilizate, and wherein said solution has a pH of from 2.5-3.5 and a concentration of said doxorubicin of from 0.1 to 100 mg/ml.

[0001] This is a continuation of U.S. Ser. No. 09/902,030, filed Jul.10, 2001 which is a continuation of U.S. Ser. No. 09/537,638, filed Mar.29, 2000, now abandoned, which is a continuation of U.S. Ser. No.09/149,381, filed Sep. 8, 1998, now U.S. Pat. No. 6,087,340, which is acontinuation of U.S. Ser. No. 08/368,402, filed Jan. 3, 1995, now U.S.Pat. No. 5,977,082, which is a continuation of U.S. Ser. No. 08/224,993,filed Apr. 8, 1994, now abandoned, which is a continuation of U.S. Ser.No. 07/827,938, filed Jan. 29, 1992, now abandoned, which is adivisional of U.S. Ser. No. 07/471,005, filed Jan. 25, 1990, now U.S.Pat. No. 5,124,318, which is a continuation of U.S. Ser. No. 07/341,249,filed Apr. 20, 1989, now abandoned, which is a continuation of U.S. Ser.No. 07/064,653, filed Jun. 22, 1987, now abandoned, which is acontinuation-in-part of U.S. Ser. No. 06/878,784, filed Jun. 26, 1986,now abandoned.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates to a stable, injectable,ready-to-use solution of an antitumor anthracycline glycoside, e.g.doxorubicin (Adriamycin®), to a process for preparing such a solution,and providing the same in a sealed container, and to a method fortreating tumors by the use of the ready-to-use solution.

[0004] 2. Description of the Related Art

[0005] The anthracycline glycoside compounds are a well known class ofcompounds in the antineoplastic group of agents, of which doxorubicin isa typical, and the most widely used, representative: Doxorubicin.Anticancer Antibiotics, Federico Arcamone, 1981, Publ: Academic Press,New York, N. Y.; Adriamycin Review, EROTC International Symposium,Brussels, May, 1974, edited by M. Staquet, Publ. Eur. Press Medikon,Ghent, Belg.; Results of Adriamycin Therapy, Adriamycin Symposium atFrankfurt/Main 1974 edited by M. Ghione, J. Fetzer and H. Maier, publ.:Springer, New York, N.Y.

[0006] In the past, solutions of anthracycline glycosides have beenprepared and the stability thereof has been studied. However, results ofthese studies have been inconsistent, and no clear parameters haveemerged for maintenance of a stable anthracycline glycoside, e.g.,doxorubicin, solution. Bosanquet, in a recent article entitled“Stability of solutions of antineoplastic agents during preparation andstorage for in vitro assays,” (Cancer Chemother. Pharmacol. 1986, 17,1-10) reviews the field of stability studies, with particular emphasison doxorubicin (Adriamycin®). He points out that “very little can becategorically stated about the stability of adriamycin, and a verycarefully designed study is urgently required to resolve theseconflicting results.”

[0007] At present, anthracycline glycoside antitumor drugs, inparticular, e.g., doxorubicin, are solely available in the form oflyophilized preparations, which need to be reconstituted beforeadministration.

[0008] Both the manufacturing and the reconstitution of suchpreparations expose the involved personnel (workers, pharmacists,medical personnel, nurses) to risks of contamination which areparticularly serious due to the toxicity of the antitumor substances.

[0009] Indeed, the Martindale Extra Pharmacopoeia 28th edition, page 175left column, reports on adverse effects of antineoplastic drugs andrecommends that “They must be handled with great care and contact withskin and eyes avoided; they should not be inhaled. Care must be taken toavoid extravasation since pain and tissue damage may ensue”.

[0010] Similarly, Scand. J. Work Environ Health vol. 10(2), pages 71-74(1984), as well as articles in Chemistry Industry, Issue Jul. 4, 1983,page 488, and Drug-Topics-Medical-Economics-Co, Issue Feb. 7, 1983, page99, report severe adverse effects observed in medical personnel exposedto use of cytostatic agents, including doxorubicin.

[0011] Even though the effect of long-term low-level exposure to suchcytotoxic drugs is not yet completely known, there is certainly a hazardfor those who regularly prepared and administer these substances in viewof the fact that they are known mutagens and carcinogens in animals andimplicated as carcinogens in man.

[0012] To administer a lyophilized preparation, double handling of thedrug is required, the lyophilized cake having to be first reconstitutedand then administered. Moreover, in some cases, the complete dissolutionof the powder may require prolonged shaking because of solubilizationproblems. Reconstitution of a lyophilized cake or powder can result information of aerosol droplets which can be inhaled or can come intocontact with skin or mucous membranes of those handling the solution.

SUMMARY OF THE INVENTION

[0013] As the risks connected with the manufacturing and thereconstitution of a lyophilized preparation would be highly reduced if aready-to-use solution of the drug were available, the present inventorshave developed a stable, therapeutically acceptable injectable solutionof an anthracycline glycoside drug, e.g. doxorubicin, whose preparationand administration does not require either lyophilization orreconstitution.

[0014] According to the present invention, there is provided a stable,injectable, sterile, pyrogen-free, anthracycline glycoside solutionwhich consists essentially of a physiologically acceptable salt of ananthracycline glycoside dissolved in a physiologically acceptablesolvent therefor, which has not been reconstituted from a lyophilizate,which has a pH of from 2.5 to 3.5 and which is preferably contained in asealed glass container.

DESCRIPTION OF THE FIGURES

[0015]FIG. 1 is a K_(obs) pH profile for doxorubicin.HCl degradation at55° C. in sterile water.

[0016]FIG. 2 is a K_(obs) pH profile for doxorubicin.HCl degradation at55° C. in 5% dextrose.

[0017]FIG. 3 is a K_(obs) pH profile for doxorubicin.HCl degradation at55° C. in 0.9% saline.

[0018]FIG. 4 is a logarithmic plot of residual doxorubicin.HClconcentration versus time.

[0019]FIG. 5 is a logarithmic plot of residual doxorubicin.HClconcentration in 0.9% NaCl.

[0020]FIG. 6 is a logarithmic plot of residual doxorubicin.HClconcentration in 5% dextrose.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] A preferred pH range for the anthracycline glycoside solution ofthis invention is from 2.5 to 3.5. A more preferred range is from 2.7 to3.5, and a particularly preferred range is from 2.7 to 3.3. A range of2.7 up to about 3.0 may also be mentioned as a useful range.

[0022] The preferred anthracycline glycoside is doxorubicinhydrochloride. It is known that doxorubicin hydrochloride is more stablein acidic than neutral or basic solutions. See Analytical Profiles ofDrug Substances, Vol. 9, Klaus Florey, ed. (Academic Press 1980). Anumber of stability studies are summarized in Bosanquet, CancerChemother. Pharmacol. 17, 1986, 1-10. However, these studies areinconsistent, in part because of the varying media used to make up thesolutions and the methods used to measure stability. Taken as a whole,the prior art has not appreciated with any degree of certainty how toprepare a stable, injectable doxorubicin solution, as a function of pH.

[0023] Martindale—The Extra Pharmacopeia 28th edition, 1828, on page205, indicates that a 0.5% solution of doxorubicin hydrochloride andwater has a pH of 3.8 to 6.5. Reconstitution of the commercial freezedried formulation which is made of doxorubicin hydrochloride andlactose, leads to a solution having a pH in the range of between 4.5 and6, containing doxorubicin at 2 milligrams per milliliter concentrationand lactose, and additionally containing sodium chloride when saline isused for reconstitution. In order to lower the pH below that ofreconstituted solutions, one must add an acid to lower the pH. In thepast there was little motivation for a user of the drug to add an acidto lower the pH, since it was not recognized that the drug was actuallymore stable at pH's between about 2.5 and 3.5. According to the presentinvention, it has been discovered that anthracycline glycosides, such asdoxorubicin hydrochloride, are stable in the pH range disclosed hereinin physiologically acceptable media. The prior art did not recognizestability for an injectable doxorubicin solution in the particularnarrow pH range disclosed herein, so this pH range was not considered tobe a particularly useful range for administration of this drug.

[0024] Preferably the solution of the invention is provided in a sealedcontainer, especially one made of glass, either in a unit dosage form orin a multiple dosage form.

[0025] In addition to doxorubicin, other anthracycline glycosidesinclude 4′-epi-doxorubicin (i.e. epirubicin), 4′-desoxy-doxorubicin(i.e. esorubicin), 4′-desoxy-4′-iodo-doxorubicin, daunorubicin and4-demethoxydaunorubicin (i.e. idarubicin).

[0026] Any physiologically acceptable salt of the anthracyclineglycoside may be used for preparing the solution of the invention.Examples of suitable salts may be, for instance, the salts with mineralinorganic acids such as hydrochloric, hydrobromic, sulfuric, phosphoric,nitric and the like, and the salts with certain organic acids such asacetic, succinic, tartaric, ascorbic, citric, glutamic, benzoic,methanesulfonic, ethanesulfonic and the like. The salt with hydrochloricacid is a particularly preferred salt, especially when the anthracyclineglycoside is doxorubicin.

[0027] Any solvent which is physiologically acceptable and which is ableto dissolve the anthracycline glycoside salt may be used. The solutionof the invention may also contain one or more additional components suchas a co-solubilizing agent (which may be the same as a solvent), atonicity adjustment agent, a stabilizing agent and a preservative.Examples of solvents, co-solubilizing agents, tonicity adjustmentagents, stabilizing agents and preservatives which can be used for thepreparation of the anthracycline glycoside solutions of the inventionare hereunder reported.

[0028] Suitable solvents and co-solubilizing agents may be, forinstance, water; physiological saline; aliphatic amides, e.g.N,N-dimethylacetamide, N-hydroxy-2-ethyl-lactamide and the like;alcohols, e.g. ethanol, benzyl alcohol and the like; glycols andpolyalcohols, e.g. propyleneglycol, glycerin and the like; esters ofpolyalcohols, e.g. diacetine, triacetine and the like; polyglycols andpolyethers, e.g. polyethyleneglycol 400, propyleneglycol methylethersand the like; dioxolanes, e.g. isopropylidenglycerin and the like;dimethylisosorbide; pyrrolidone derivatives, e.g. 2-pyrrolidone,N-methyl-2-pyrrolidone, polyvinylpyrrolidone (co-solubilizing agentonly) and the like; polyoxyethylenated fatty alcohols, e.g., Brij® andthe like; esters of polyoxyethylenated fatty acids, e.g., Cremophor®,Myrj® and the like; polysorbates, e.g., Tweens®, polyoxyethylenederivatives of polypropyleneglycols, e.g., Pluronics®.

[0029] A particularly preferred co-solubilizing agent ispolyvinylpyrrolidone.

[0030] Suitable tonicity adjustment agents may be, for instance,physiologically acceptable inorganic chlorides, e.g. sodium chloride;dextrose; lactose; mannitol; sorbitol and the like.

[0031] Preservatives suitable for physiological administration may be,for instance, esters of parahydroxybenzoic acid (e.g., methyl, ethyl,propyl and butyl esters, or mixtures of them), chlorocresol and thelike.

[0032] Suitable stabilizing agents include monosaccharides (e.g.,galactose, fructose, and fucose), disaccharides (e.g., lactose),polysaccharides (e.g., dextran), cyclic oligosaccharides (e.g., α-, β-,γ-cyclodextrin), aliphatic polyols (e.g., mannitol, sorbitol, andthioglycerol), cyclic polyols (e.g. inositol) and organic solvents(e.g., ethyl alcohol and glycerol). These may be included inconcentrations of from about 0.25-10% w/v, preferably 0.5-5% w/v in thesolution.

[0033] The above mentioned solvents and co-solubilizing agents, tonicityadjustment agents, stabilizing agents and preservatives can be usedalone or as a mixture of two or more of them.

[0034] Examples of preferred solvents are water, ethanol, polyethyleneglycol and dimethylacetamide as well as mixtures in various proportionsof these solvents. Water is a particularly preferred solvent. Othersolvents giving good results in terms of stability are 0.9% sodiumchloride solution (i.e., physiological saline), and, especially, 5%dextrose solution, 5% mannitol solution and 5% sorbitol solution, i.e.,aqueous solutions containing approximately 5% of, respectively,dextrose, mannitol or sorbitol. Small variations (±2-3%) of theseadditional ingredients also fall within the scope of the presentinvention.

[0035] To adjust the pH within the range of from 2.5 to about 3.5, aphysiologically acceptable acid is added to the solution of theanthracycline glycoside. The acid should be a physiologically acceptableacid, e.g., an inorganic mineral acid such as hydrochloric, hydrobromic,sulfuric, phosphoric, nitric and the like, or an organic acid such asacetic, succinic, tartaric, ascorbic, citric, glutamic, benzoic,methanesulphonic, ethanesulfonic and the like, or also an acidicphysiologically acceptable buffer solution, e.g., a chloride buffer, anacetate buffer, a phosphate buffer and the like.

[0036] In the solutions of the invention the concentration of theanthracycline glycoside may vary within broad ranges, preferably from0.1 mg/ml to 100 mg/ml, in particular from 0.1 mg/ml to 50 mg/ml, mostpreferably from 1 mg/ml to 20 mg/ml.

[0037] The preferred ranges of concentration may be slightly differentfor different anthracycline glycosides. Thus, for example, preferredconcentrations for doxorubicin are from about 2 mg/ml to about 50 mg/ml,preferably from 2 mg/ml to 20 mg/ml, particularly appropriate valuesbeing 2 mg/ml and 5 mg/ml. Similar concentrations are preferred also for4′-epi-doxorubicin, 4′-desoxy-doxorubicin and4′-desoxy-4′-iodo-doxorubicin. Preferred ranges of concentration fordaunorubicin and 4-demethoxy-daunorubicin are from 0.1 mg/ml to 50mg/ml, preferably from 1 mg/ml to 20 mg/ml, concentrations of 1 mg/mland 5 mg/ml being particularly appropriate.

[0038] Suitable packaging for the anthracycline glycoside solutions maybe all approved containers intended for parenteral use, such as plasticand glass containers, ready-to-use syringes and the like. Preferably thecontainer is a sealed glass container, e.g. a vial or an ampoule. Ahermetically sealed glass vial is particularly preferred.

[0039] According to a particularly preferred feature of the invention,there is provided, in a sealed glass container, a sterile, pyrogen-free,injectable doxorubicin solution which consists essentially of aphysiologically acceptable salt of doxorubicin dissolved in aphysiologically acceptable solvent therefor, which has not beenreconstituted from a lyophilizate and which has a pH of from 2.5 to 3.5.

[0040] In the above indicated preferred feature of the invention thephysiologically acceptable salt of doxorubicin may be, e.g. the saltwith a mineral inorganic acid such as hydrochloric, hydrobromic,sulfuric, phosphoric, nitric and the like, or the salt with an organicacid such as acetic, succinic, tartaric, ascorbic, citric, glutamic,benzoic, methanesulfonic, ethanesulfonic and the like. The hydrochloridesalt is a particularly preferred salt.

[0041] For the solution hereabove indicated as a preferred feature ofthe invention suitable solvents, co-solubilizing agents, tonicityadjustment agents, stabilizing agents and preservatives may be the sameas those previously recited in this specification. Water is aparticularly preferred solvent.

[0042] Also, the physiologically acceptable acid which is added toadjust the pH to from 2.5 to about 3.5 may be one of those previouslyspecified. When it is desired to adjust the pH of the above saidpreferred solution to a value of from 2.5 to about 3.5, hydrochloricacid is an especially preferred acid. Preferred pH values for the abovesaid preferred solutions of the invention are from about 2.7 to about3.3.

[0043] Though the concentration of doxorubicin in the above preferredfeature may vary within the broad range from 0.1 mg/ml to 100 mg/ml,preferred concentrations are from 2 mg/ml to 50 mg/ml, most preferablyfrom 2 mg/ml to 20 mg/ml: examples of especially preferredconcentrations of doxorubicin are 2 mg/ml and 5 mg/ml.

[0044] The invention also provides a process for producing a sterile,pyrogen-free anthracycline glycoside solution with a pH of from 2.5 to3.5 which process comprises dissolving a physiologically acceptable saltof the anthracycline glycoside, which salt is not in the form of alyophilizate, in a physiologically acceptable solvent therefor; adding aphysiologically acceptable acid or buffer to adjust the pH within thesaid range as desired; and passing the resulting solution through asterilizing filter.

[0045] One or more additional components such as co-solubilizing agents,tonicity adjustment agents, stabilizing agents and preservatives, forinstance of the kind previously specified, may be added to the solutionprior to passing the solution through the sterilizing filter.

[0046] With the solutions of the invention it is possible to obtaincompositions having a very high concentration of the anthracyclineglycoside active substance even at 50 mg/ml and more. This constitutes agreat advantage over the presently available lyophilized preparationswherein high concentrations of anthracycline glycoside can only beobtained with difficulty because of solubilization problems encounteredin reconstitution, mainly with saline. The presence of the excipient,e.g. lactose, in the lyophilized cake, and its generally high proportionin respect of the active substance, even up to 5 parts of excipient perpart of active substance, has a negative effect on solubilization sothat difficulties may arise in obtaining dissolution of the lyophilizedcake, especially for concentrations of anthracycline glycoside higherthan 2 mg/ml.

[0047] The solution of the invention are characterized by a goodstability. Solutions in various solvents and with different pH's andconcentrations have been found to be stable for long periods attemperatures accepted for the storage of pharmaceutical preparations.This is illustrated in the Examples which follow.

[0048] Due to the well known anti-tumor activity of the anthracyclineglycoside active drug substance, the pharmaceutical compositions of theinvention are useful for treating tumors in both human and animal hosts.Examples of tumors that can be treated are, for instance, sarcomas,including osteogenic and soft tissue sarcomas, carcinomas, e.g.,breast-, lung-, bladder-, thyroid-, prostate- and ovarian carcinoma,lymphomas, including Hodgkin and non-Hodgkin lymphomas, neuroblastoma,melanoma, myeloma, Wilms tumor, and leukemias, including acutelymphoblastic leukemia and acute myeloblastic leukemia. SeeKirk-Othmer's Encyclopedia of Chemical Technology, Volume 5, pages478-479 (1979).

[0049] Examples of specific tumors that can be treated are MoloneySarcoma Virus, Sarcoma 180 Ascites, solid Sarcoma 180, grosstransplantable leukemia, L 1210 leukemia and lymphocytic P 388 leukemia.

[0050] Thus, according to the invention there is also provided a methodof inhibiting the growth of a tumor, in particular one of thoseindicated above, which comprises administering to a host suffering fromsaid tumor an injectable solution according to the invention containingthe active drug substance in an amount sufficient to inhibit the growthof said tumor.

[0051] The injectable solutions of the invention are administered byrapid intravenous injection or infusion according to a variety ofpossible dose schedules. A suitable dose schedule for doxorubicin maybe, for example, of 60 to 75 mg of active drug substance per m² of bodysurface given as a single rapid infusion and repeated at 21 days; analternative schedule may be of 30 mg/m² per day by intravenous route for3 days, every 28 days. Suitable dosages for 4′-epi-doxorubicin and4′-desoxy-doxorubicin may be, for instance, of 75 to 90 mg/m² given in asingle infusion to be repeated at 21 days, and similar dosages may beuseful also for 4′-desoxy-4′-iodo-doxorubicin.

[0052] Idarubicin, i.e. 4-demethoxy-daunorubicin, may be administeredintravenously at a single dose of 13-15 mg/m² every 21 days in thetreatment of solid tumors, while in the treatment of leukemias apreferred dose schedule is, e.g., of 10-12 mg/m² day be intravenousroute for 3 days, to be repeated every 15-21 days; similar dosages maybe followed also for daunorubicin.

EXAMPLES

[0053] The invention now being generally described, the same will bebetter understood by reference to certain specific examples which areincluded herein for purposes of illustration only and are not intendedto be limiting of the invention or any embodiment thereof, unlessspecified.

[0054] With reference to the first three examples, the stabilitycontrols on the ready-to-use solutions were carried out by means of highperformance liquid chromatography (HPLC), at the following experimentalconditions:

[0055] Liquid chromatograph: Varian model 5010

[0056] Spectrophotometric detector: Knauer model 8700

[0057] Integrating recorder: Varian model CDS 401

[0058] Injection valve: Rheodyne model 7125 fitted with a 10 mcl sampleloop

[0059] Chromatographic column: Waters .mu.-Bondapak C18 (length=300 mm;inner diameter=3.9 mm; average particle size=10 mcm)

[0060] Column temperature: ambient (about 22° C.±2° C.)

[0061] Mobile phase: water:acetonitrile (69:31 v/v) adjusted to pH 2with phosphoric acid, filtered (sintered glass filter, 1 mcm or finerporosity) and deaerated

[0062] Mobile phase flow rate: 1.5 ml/min

[0063] Analytical wavelength: 254±1 nm

[0064] Integrating recorder sensitivity: 512

[0065] Chart speed: 1 cm/min

[0066] At these conditions, the peak of the anthracycline glycosideshowed a retention time of about 6 minutes.

[0067] The obtained results are reported in Tables accompanying Examples1-3.

[0068] The extrapolation of the analytical data in order to determinethe time when the 90% of the initial assay could be expected (t₉₀ value)was made following an Arrhenius plot.

[0069] This procedure of analytical data treatment is well known andwidely used and described in the art: see e.g., Chemical Stability ofPharmaceuticals, Kennet A. Connors, Gordon L. Amidon, Lloyd Kennon,Publ. John Wiley and Sons, New York, N.Y., 1979.

[0070] The term “teflon” recurring in the examples refers to “Teflon™”.

Example 1

[0071] Composition for 80 vials (for 1 vial) Doxorubicin·HCl 0.8 g (10mg) Water for injections 0.4 l (5 ml) q.s. to

[0072] Doxorubicin.HCl (0.80 g) was dissolved in 90 percent of theamount of water for injections, de-aerated by nitrogen bubbling. The pHof the solution was not adjusted. Further de-aerated water forinjections was then added to bring the solution to its final volume(0.40 l).

[0073] The solution was filtered through a 0.22 μm microporous membraneunder nitrogen pressure. Volumes of 5 ml of the solution weredistributed into type I-colorless glass vials having 5/7 ml capacity.The vials were then closed with chlorobutyl teflon-faced rubber stoppersand sealed with aluminum caps.

[0074] The stability of the solutions in the vials was tested. The vialswere stored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 12 weeks (4° C.).

[0075] The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 1: TABLE 1 INITIAL VALUES Concentration: 1.994 mg/mlpH = 5.2 Relative % Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C.TIME Conc. Rel. % Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/mlAssay mg/ml Assay mg/ml Assay mg/ml Assay 1 1.992 99.9 1.917 96.1 1.76888.7 1.493 75.0 2 1.843 92.4 1.618 81.1 1.166 58.5 3 1.774 89.0 1.50675.5 0.830 41.6 4 1.974 99.0 1.720 86.3 1.393 69.9 12 1.980 99.3

[0076] Similar stability data can be observed also for analogoussolutions containing either doxorubicin hydrochloride at 5 mg/mlconcentration, or 4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-desoxy-4′-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

Example 2

[0077] Composition for 80 vials (for 1 vial) Doxorubicin·HCl 0.8 g (10mg) Hydrochloric acid 0.1 N pH = 3 (pH = 3) q.s. to Water for injections0.4 l (5 ml) q.s. to

[0078] Doxorubicin.HCl (0.8 g) was dissolved in 90 percent of the amountof water for injections, de-aerated by nitrogen bubbling. Thehydrochloric acid was then added dropwise to adjust the pH of thesolution to 3. Further de-aerated water for injections was then added tobring the solution to its final volume (0.4 l).

[0079] The solution was filtered through a 0.22 μm microporous membraneunder nitrogen pressure. Volumes of 5 ml of the solution weredistributed into type I-colorless glass vials having 5/7 ml capacity.The vials were then closed with chlorobutyl teflon-faced rubber stoppersand sealed with aluminum caps.

[0080] The stability of the solutions in the vials was tested. The vialswere stored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 12 weeks (4° C.).

[0081] The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 2: TABLE 2 INITIAL VALUES Concentration: 1.992 mg/mlpH = 3.0 Relative % Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C.TIME Conc. Rel. % Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/mlAssay mg/ml Assay mg/ml Assay mg/ml Assay 1 1.995 100.2 1.952 98.0 1.91996.3 1.493 75.0 2 1.889 94.8 1.851 92.9 1.036 51.9 3 1.876 94.2 1.56578.6 0.730 36.7 4 1.979 99.4 1.808 90.8 1.393 69.9 12 1.972 99.0

[0082] Similar stability data can be observed also for analogoussolutions containing either doxorubicin hydrochloride at 5 mg/mlconcentration, or 4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-desoxy-4′-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

Example 3

[0083] Composition for 80 vials (for 1 vial) Doxorubicin·HCl 0.8 g (10mg) Hydrochloric acid 0.1 N pH = 3 (pH = 3) q.s. to Water for injections0.4 l (5 ml) q.s. to

[0084] Doxorubicin.HCl (8.0 g) was dissolved in 90 percent of the amountof water for injections, de-aerated by nitrogen bubbling. Thehydrochloric acid was then added dropwise to adjust the pH of thesolution 3. Further de-aerated water for injections was then added tobring the solution to its final volume (0.4 l).

[0085] The solution was filtered through a 0.22 μm microporous membraneunder nitrogen pressure. Volumes of 5 ml of the solution weredistributed into type I-colorless glass vials having 5/7 ml capacity.The vials were then closed with chlorobutyl teflon-faced rubber stoppersand sealed with aluminum caps.

[0086] The stability of the solutions in the vials was tested. The vialswere stored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 12 weeks (4° C.).

[0087] The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 3: TABLE 3 INITIAL VALUES Concentration: 20.06 mg/mlpH = 2.95 Relative % Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C.TIME Conc. Rel. % Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/mlAssay mg/ml Assay mg/ml Assay mg/ml Assay 1 20.06 100.0 19.56 97.5 17.8488.9 12.31 61.4 2 18.87 94.1 15.61 77.8 7.09 35.3 3 18.24 90.9 13.4166.8 3.13 15.6 4 19.91 99.2 17.51 87.3 11.07 55.2 12 19.80 98.7

[0088] Similar stability data can be observed for analogous solutionscontaining 4′-epi-doxorubicin or 4′-desoxy-doxorubicin, as hydrochloridesalts, at the same 20 mg/ml concentration.

[0089] The following examples regarding stability profile and shelf-lifeforecasts were carried out under accelerated temperature conditions on5.0 ml of 2 mg/ml doxorubicin.HCl solutions in a container-closuresystem consisting of: glass type I, 8 ml top capacity vial; teflon-facedchlorobutyl rubber bung; aluminum seal.

[0090] pH-Stability Profile at 55° C. of Doxorubicin.HCl Solutions inSterile Water 5% Dextrose, 0.9% Saline

[0091] 2 mg/ml doxorubicin.HCl solutions were prepared in the following1=0.05 buffers: a) glycine.HCl pH 2.0, 2.5 and 3.0; b) formate pH 3.5;c) acetate pH 4.0, 5.0 and 5.5.

[0092] 5.0 ml of each solution in glass vials were stored at 55° C. andanalyzed at prefixed times (up to 120 hours) for doxorubicin.HCl assayand pH.

[0093] Tables 4, 5 and 6 give the doxorubicin.HCl residual concentrationand percent stability at 55° C., at different pH's and times of storagefor sterile water, 5% dextrose and 0.9% saline solutions, respectively.

[0094] The doxorubicin.HCl assays are the mean of three independentdeterminations performed by the USP HPLC method (USP XXI). At each pHvalue, the pseudo-first order rate constants (K_(obs)) for thedegradation were calculated by linear regression analysis of the naturallogarithm of the residual concentration of doxorubicin.HCl (|Dx|_(t))versus time as depicted by the following equation:

ln |Dx| _(t)=ln |Dx| _(o) −K _(obs) ^(·) t

[0095] J. Thuro Carstensen, Theory of Pharmaceutical Systems, Volume1/General Principles, page 172, Academic Press, New York and London 1972

[0096] Kenneth A. Connors, Gordon L. Amidon, Lloyd Kennon, ChemicalStability of Pharmaceuticals, chapter 2, John Wiley and Sons, New York1979

[0097] Arthur Osol, Remington's Pharmaceutical Sciences, 16th Edition,chapter 18, Mack Publishing Company, Easton, Pa. 1980

[0098] Valentino J. Stella, Chemical and Physical Bases Determining theInstability and Incompatibility of Formulated Injectable Drugs, Journalof Parenteral Sciences & Technology, July-August 1986, page 142.

[0099] Tables 7, 8 and 9 give the observed rate constants (K_(obs)) forthe degradation kinetics of doxorubicin.HCl at 55° C. and at differentpH's for sterile water, 5% dextrose and 0.9% saline solutions,respectively.

[0100]FIGS. 1, 2, and 3 show the K_(obs)-pH profile for thedoxorubicin.HCl degradation at 55° C. in the above mentioned media. Thedata in Tables 4-9 and the FIGS. 1-3 evidence that the 2 mg/mldoxorubicin.HCl solutions show at 55° C. the maximum stability in the pHrange about 3.0-3.5 (±0.2, e.g., 2.8, 3.2 and 3.3, 3.7) for all thethree media tested. The range of from 2.5 to 3.0 is also a range ofnotable stability.

[0101] A common behavior as to stability is thus evidenced for aqueoussolutions in general, since no practical differences in stability areobserved when going from sterile water as such to sterile watercontaining a tonicity adjustment agent, either ionic, such as, e.g.,sodium chloride, or non-ionic, such as, e.g., dextrose. TABLE 4Accelerated (55° C.) stability data of 2 mg/ml doxorubicin·HCl solutionsin sterile water at various pHs Time (hours) Buffers Tests 0 8 16 24 4872 120 pH 2.0 Doxorubicin·HCl 2.022 1.892 1.669 1.554 1.145 0.801glycine-HCl assay · mg/ml · % stability 100.0 93.6 82.6 76.9 56.6 39.6pH 2.00 2.01 2.02 2.01 2.01 2.02 pH 2.5 Doxorubicin·HCl assay 1.9921.926 1.835 1.718 1.557 1.00 glycine-HCl mg/ml % stability 100.0 96.792.1 86.2 78.2 50.2 pH 2.51 2.50 2.50 2.52 2.51 2.52 pH 3.0Doxorubicin·HCl assay 2.003 1.958 1.881 1.831 1.696 1.525 1.258glycine-HCl mg/ml % stability 100.0 97.8 93.9 91.4 84.7 76.1 62.8 pH3.00 3.03 3.02 3.02 3.01 3.02 3.00 pH 3.5 Doxorubicin·HCl assay 2.0351.950 1.887 1.840 1.650 1.538 1.241 formate mg/ml % stability 100.0 95.892.7 90.4 81.1 75.6 61.0 pH 3.51 3.51 3.51 3.51 3.52 3.52 3.51 pH 4.0Doxorubicin·HCl assay 2.032 1.788 1.681 1.561 1.167 acetate mg/ml %stability 100.0 88.0 82.7 76.8 57.4 pH 4.00 4.00 4.04 4.02 4.02 pH 5.0Doxorubicin·HCl assay 2.019 1.823 1.688 1.512 1.060 acetate mg/mlstability 100.0 90.3 83.6 74.9 52.5 pH 5.03 5.05 5.04 5.04 5.05 pH 5.5Doxorubicin·HCl assay 2.047 1.808 1.427 1.228 0.903 acetate mg/ml %stability 100.0 88.3 69.7 60.0 44.1 pH 5.50 5.53 5.53 5.54 5.56

[0102] TABLE 5 Accelerated (55° C.) stability data of 2 mg/mldoxorubicin·HCl solutions in 5% dextrose at various pHs Time (hours)Buffers Tests 0 8 16 24 34 48 72 96 120 pH 2.0 Doxorubicin·HCl assay1.993 1.851 1.683 1.513 1.361 1.078 0.765 glycine-HCl mg/ml % stability100.0 92.8 84.4 75.9 68.3 54.1 38.4 pH 2.14 2.13 2.14 2.15 2.18 2.212.16 pH 2.5 Doxorubicin·HCl assay 1.967 1.897 1.822 1.760 1.682 1.4991.305 glycine-HCl mg/ml % stability 100.0 96.4 92.6 89.5 85.5 76.2 66.3pH 2.56 2.56 2.56 2.58 2.60 2.56 2.61 pH 3.0 Doxorubicin·HCl assay 1.9751.908 1.832 1.645 1.508 1.344 1.206 glycine-HCl mg/ml % stability 100.096.6 92.7 83.3 76.4 68.0 61.1 pH 3.04 3.05 3.05 3.06 3.00 3.13 3.10 pH3.5 Doxorubicin·HCl assay 1.983 1.897 1.858 1.622 1.324 1.222 formatemg/ml % stability 100.0 95.7 93.7 81.8 66.8 61.6 pH 3.58 3.59 3.60 3.633.60 3.63 pH 4.0 Doxorubicin·HCl assay 2.003 1.913 7.716 1.665 1.4871.312 1.081 acetate mg/ml % stability 100.0 95.5 85.6 83.1 74.2 65.553.9 pH 4.10 4.10 4.11 4.11 4.16 4.15 4.12 pH 5.0 Doxorubicin·HCl assay2.012 1.906 1.673 1.608 1.416 1.163 acetate mg/ml % stability 100.0 94.783.2 79.9 70.4 57.8 pH 5.06 5.06 5.06 5.06 5.07 5.04 pH 5.5Doxorubicin·HCl assay 1.991 1.841 1.470 1.246 1.091 acetate mg/ml %stability 100.0 92.5 73.8 62.6 54.8 pH 5.56 5.54 5.48 5.50 5.46

[0103] TABLE 6 Accelerated (55° C.) stability data of 2 mg/mldoxorubicin·HCl solutions in 0.9% saline at various pHs Time (hours)Buffers Tests 0 4 8 16 24 34 48 72 96 120 pH 2.0 Doxorubicin·HCl assay1.998 1.857 1.580 1.397 1.231 0.931 0.701 glycine-HCl mg/ml % stability100.0 92.9 79.1 69.9 61.6 46.6 35.1 pH 2.16 2.16 2.18 2.16 2.22 2.202.19 pH 2.5 Doxorubicin·HCl assay 1.946 1.875 1.670 1.602 1.368 1.132glycine-HCl mg/ml % stability 100.0 96.3 85.8 82.3 70.3 58.1 pH 2.592.59 2.59 2.58 2.62 2.62 pH 3.0 Doxorubicin·HCl assay 1.994 1.818 1.7711.571 1.375 1.205 1.003 glycine-HCl mg/ml % stability 100.0 91.2 88.878.8 69.0 60.4 50.3 pH 3.06 3.07 3.07 3.08 3.13 3.14 3.12 pH 3.5Doxorubicin·HCl assay 1.997 1.824 1.742 1.543 1.323 1.176 0.919 formatemg/ml % stability 100.0 91.4 87.2 77.3 66.2 58.9 46.0 pH 3.58 3.56 3.563.66 3.61 3.64 3.63 pH 4.0 Doxorubicin·HCl assay 1.972 1.885 1.828 1.6531.594 acetate mg/ml % stability 100.0 95.6 92.7 83.8 80.8 pH 4.10 4.104.10 4.10 4.11 pH 5.0 Doxorubicin·HCl assay 1.979 1.732 1.469 1.4421.278 acetate mg/ml % stability 100.0 87.5 74.2 72.8 64.6 pH 5.04 5.065.04 5.05 5.05 pH 5.5 Doxorubicin·HCl assay 2.023 1.847 1.548 1.330acetate mg/ml % stability 100.0 91.3 76.5 65.7 pH 5.58 5.56 5.55 5.53

[0104] TABLE 7 K_(obs) values (1/days) for the degradation ofdoxorubicin·HCl 2 mg/ml solutions in sterile water at various pHs at 55°C. Buffer pH K_(obs) × 10³ 95% confidence limits Glycine-HCl 2.0 309.5±12.6 (I = 0.05) Glycine-HCl 2.5 138.3 ±0.6 (I = 0.05) Glycine-HCl 3.093.1 ±4.6 (I = 0.05) Formate 3.5 96.7 ±4.4 (I = 0.05) Acetate 4.0 269.8±18.7 (I = 0.05) Acetate 5.0 322.6 ±19.2 (I = 0.05) Acetate 5.5 415.4±45.7 (I = 0.05)

[0105] TABLE 8 K_(obs) values (1/days) for the degradation ofdoxorubicin·HCl 2 mg/ml solutions in 5% dextrose at various pHs at 55°C. Buffer pH K_(obs) × 10³ 95% confidence limits Glycine-HCl 2.0 323.8±17.2 (I = 0.05) Glycine-HCl 2.5 138.7 ±9.9 (I = 0.05) Glycine-HCl 3.0100.5 ±5.9 (I = 0.05) Formate 3.5 132.0 ±20.7 (I = 0.05) Acetate 4.0209.7 ±12.7 (I = 0.05) Acetate 5.0 273.1 ±27.7 (I = 0.05) Acetate 5.5453.7 ±59.2 (I = 0.05)

[0106] TABLE 9 K_(obs) values (1/days) for the degradation ofdoxorubicin·HCl 2 mg/ml solutions in 0.9% saline at various pHs at 55°C. Buffer pH K_(obs) × 10³ 95% confidence limits Glycine-HCl 2.0 362.4±19.4 (I = 0.05) Glycine-HCl 2.5 276.5 ±30.2 (I = 0.05) Glycine-HCl 3.0133.2 ±8.0 (I = 0.05) Formate 3.5 148.1 ±11.1 (I = 0.05) Acetate 4.0215.7 ±35.4 (I = 0.05) Acetate 5.0 301.2 ±60.1 (I = 0.05) Acetate 5.5430.3 ±69.9 (I = 0.05)

[0107] Shelf-Life (t 90%) Forecast of Doxorubicin.HCl 2 mg/ml SterileWater Solution Adjusted to pH 3.0

[0108] 5.0 ml of doxorubicin.HCl 2 mg/ml aqueous solution adjusted to pH3.0 with 0.5 N HCl were stored, in glass vials, at:

[0109] a) 55° C. for 21 days, b) 45° C. and 35° C. for 28 days, c) 27°C. for 90 days.

[0110] b) At prefixed times the vials were analyzed for doxorubicin.HClassay and pH.

[0111] The logarithmic plots of the residual concentration versus timewere linear and indicated the degradation of the drug to followpseudo-first order kinetics at constant pH and temperature.

[0112] The observed rate constants (K_(obs)) for the degradation werecalculated again by linear regression analysis of a plot of the naturallogarithm of the residual concentration of doxorubicin.HCl (|Dx|_(t))versus time as depicted by the equation previously reported:

ln |Dx| _(t)=ln |Dx| _(o) −K _(obs) ^(·) t

[0113] The Arrhenius equation for the degradation process was calculatedfrom the K_(obs) obtained from the different temperatures taken inaccount for the testing (table 11).

[0114] Applying the equation, the rate constants for the pseudo-firstorder reactions at 4° C., 8° C., 15° C. and 27° C. were calculated,together with the expected t_(90%) at these temperatures.

[0115] Table 10 gives the doxorubicin.HCl residual concentration andpercent stability at pH 3.0, at different temperatures and times ofstorage.

[0116]FIG. 4 gives the logarithm of the remaining doxorubicin.HClconcentration versus time at different temperatures.

[0117] The t.₉₀% forecasts (table 11) show that a commerciallymeaningful shelf-life can be attributed to doxorubicin.HCl 2 mg/ml pH3.0 aqueous solution only if the product is stored in a refrigerator(between 2° C. and 8° C.). TABLE 10 Accelerated stability data ofdoxorubicin·HCl 2 mg/ml pH 3.0 solutions in sterile water at differenttimes and temperatures Storage Time (days) Temperature Tests 0 4 8 14 3228 60 90 27° C. Doxorubicin·HCl assay 1.992 1.993 1.988 1.962 1.9411.908 1.850 mg/ml % stability 100.0 100.1 99.8 98.5 97.4 95.8 92.9 pH3.00 2.95 2.94 2.95 2.94 2.96 2.93 35° C. Doxorubicin·HCl assay 1.9921.985 1.952 1.889 1.876 1.808 mg/ml % stability 100.0 99.6 98.0 94.894.2 90.8 pH 3.00 2.96 2.98 2.93 2.92 2.92 45° C. Doxorubicin·HCl assay1.992 1.919 1.851 1.677 1.565 1.393 mg/ml % stability 100.0 96.3 92.984.2 78.6 69.9 pH 3.00 2.97 2.95 2.85 2.92 2.90 55° C. Doxorubicin·HClassay 1.992 1.760 1.493 1.036 0.730 mg/ml % stability 100.0 88.4 74.952.0 36.6 pH 3.00 2.94 2.90 2.80 2.82

[0118] TABLE 11 Arrhenius approach. Pseudo-first order rate constants,Arrhenius equation, calculated t_(90%) PSEUDO-FIRST ORDER RATECONSTANTS, OBSERVED VALUES (K_(obs)) Temperature K_(obs) × 10³ (1/days)Correlation Coefficient 27° C. 0.850 0.986 35° C. 3.506 0.983 45° C.12.790 0.995 55° C. 49.340 0.995

[0119] Arrhenius Equation from 27° C., 35° C., 45° C., and 55° C. RateConstants

[0120] In K_(obs)=−14083/T+39.95

[0121] correlation coefficient=0.9988

[0122] Pseudo-First Order Rate Constants, Calculated Values (K)Temperature K × · 10³ (1/days) t_(90%) (days) 95% confidence limits  4°C. 0.019 5,652  3,079-10,380  8° C. 0.038 2,745 1,603-4,697 15° C. 0.130810   532-1,238 27° C. 0.918 115  89-147

[0123] Shelf-Life (t 90%) Forecast of Doxorubicin.HCl 2 mg/ml 0.9%Sodium Chloride Solution Adjusted to pH 3.0

[0124] 5.0 ml of doxorubicin.HCl 2 mg/ml solution in 0.9% sodiumchloride adjusted to pH 3.0 with 0.5 N HCl were stored, in glass vials,at: a) 60° C. for 4 days, b) 55° C. for 14 days, c) 45° C. for 21 days,d) 35° C. for 28 days.

[0125] At prefixed times the vials were analyzed for doxorubicin.HClassay and pH.

[0126] The logarithmic plots of the residual concentration versus timewere linear and indicated the degradation of the drug to followpseudo-first order kinetics at constant pH and temperature.

[0127] The observed rate constants (K_(obs)) for the degradation werecalculated again by linear regression analysis of a plot of the naturallogarithm of the residual concentration of doxorubicin.HCl (|Dx|_(t))versus time as depicted by the equation previously reported:

ln |Dx| _(t)=ln |Dx| _(o) −K _(obs) ^(·) t

[0128] The Arrhenius equation for the degradation process was calculatedfrom the K_(obs) obtained from the different temperatures taken inaccount for the testing (table 13).

[0129] Applying the equation, the rate constants for the pseudo-firstorder reactions at 4° C., 8° C., 15° C. and 27° C. were calculated,together with the expected t.₉₀% at these temperatures.

[0130] Table 12 gives the doxorubicin.HCl residual concentration andpercent stability at pH 3.0, at different temperatures and times ofstorage.

[0131]FIG. 5 gives the logarithm of the remaining doxorubicin.HClconcentration versus time at different temperatures. TABLE 12Accelerated stability data of doxorubicin·HCl 2 mg/ml in 0.9% sodiumchloride at different times and temperatures Storage Time (days)Conditions Tests 0 1 2 3 4 8 11 14 21 28 35° C. Doxorubicin·HCl assay2.061 2.045 1.946 1.932 1.852 mg/ml % stability 100.0 99.2 94.4 93.789.9 pH 3.05 2.98 2.92 2.92 2.98 45° C. Doxorubicin·HCl assay 2.0611.996 1.724 1.517 1.344 mg/ml % stability 100.0 96.5 83.6 73.6 65.2 pH3.05 2.98 2.97 2.98 2.93 55° C. Doxorubicin·HCl assay 2.061 1.450 1.0660.900 mg/ml % stability 100.0 70.4 51.7 43.7 pH 3.05 2.90 2.97 2.95 60°C. Doxorubicin·HCl assay 2.061 1.742 1.481 1.290 1.050 mg/ml % stability100.0 84.5 71.9 62.6 50.9 pH 3.05 2.97 2.96 2.98 2.96

[0132] The t_(90%) forecasts (table 13) show that a commerciallymeaningful shelf-life can be attributed to doxorubicin.HCl 2 mg/ml pH3.0 solution in 0.9% sodium chloride only if the product is stored in arefrigerator (between 2° C. and 8° C.).

[0133] Table 13—Doxorubicin.HCl 2 mg/ml pH 3.0 Solution in 0.9% NaCl

[0134] Arrhenius approach. Pseudo-first order rate constants,

[0135] Arrhenius equation, calculated t_(90%) PSEUDO-FIRST ORDER RATECONSTANTS, OBSERVED VALUES (K_(obs)) Temperature K_(obs) × 10³ (1/days)Correlation Coefficient 35° C. 3.89 0.965 45° C. 21.61 0.987 55° C.75.90 0.996 60° C. 164.90 0.998

[0136] Arrhenius Equation from 35° C., 45° C., 55° C. and 60° C. RateConstants

ln K _(obs)=−15100/T+43.53

[0137] correlation coefficient=0.9986 PSEUDO-FIRST ORDER RATE CONSTANTS,CALCULATED VALUES (K) Temperature K × 10³ (1/days) t_(90%) (days) 95%confidence limits  4° C. 0.017 6,166 1,670-22,756  8° C. 0.037 2,838 861-9,351 15° C. 0.137 768  281-2,105 27° C. 1.112 94 45-197

[0138] Shelf-Life (t 90%) Forecast of Doxorubicin.HCl 2 mg/ml Solutionin 5% Dextrose Adjusted to pH 3.0

[0139] 5.0 ml of doxorubicin.HCl 2 mg/ml solution in 5% dextroseadjusted to pH 3.0 with 0.5 N HCl were stored, in glass vials, at: a)60° C. for 8 days, b) 55° C. for 17 days, c) 45° C. and 35° C. for 28days.

[0140] At prefixed times the vials were analyzed for doxorubicin.HClassay and pH.

[0141] The logarithmic plots of the residual concentration versus timewere linear and indicated the degradation of the drug to followpseudo-first order kinetics at constant pH and temperature. The observedrate constants (K_(obs)) for the degradation was calculated again bylinear regression analysis of a plot of the natural logarithm of theresidual concentration of doxorubicin.HCl (|Dx|_(t)) versus time asdepicted by the equation previously reported:

ln |Dx| _(t)=ln |Dx| _(o) ^(−K) _(obs) ^(·) t

[0142] The Arrhenius equation for the degradation process was calculatedfrom the K_(obs) obtained from the different temperatures taken inaccount for the testing (table 15).

[0143] Applying the equation, the rate constants for the pseudo-firstorder reactions at 4° C., 8° C., and 15° C. and 27° C. were calculated,together with the expected t_(90%) at these temperatures.

[0144] Table 14 gives the doxorubicin.HCl residual concentration andpercent stability at pH 3.0, at different temperatures and times ofstorage.

[0145]FIG. 6 gives the logarithm of the remaining doxorubicin.HClconcentration versus time at different temperatures.

[0146] The t_(90%) forecasts (table 15) show that a commerciallymeaningful shelf-life can be attributed to doxorubicin.HCl 2 mg/ml pH3.0 5% dextrose solution only if the product is stored in a refrigerator(between 2° C. and 8° C.). TABLE 14 Accelerated stability data ofdoxorubicin·HCl 2 mg/ml pH 3.0 solution in 5% dextrose at differenttimes and temperatures Storage Time (days) Conditions Tests 0 2 4 6 8 1114 17 21 28 35° C. Doxorubicin·HCl assay 2.114 2.044 2.034 2.015 1.9341.859 mg/ml % stability 100.0 96.7 96.2 53.3 91.5 87.9 pH 3.02 2.98 2.942.95 2.90 2.94 45° C. Doxorubicin·HCl assay 2.114 1.940 1.870 1.6841.510 1.410 mg/ml % stability 100.0 91.8 88.5 79.7 71.5 66.7 pH 3.022.97 2.98 2.95 2.96 2.96 55° C. Doxorubicin·HCl assay 2.114 1.718 1.4151.112 0.957 0.796 mg/ml % stability 100.0 81.3 66.9 52.6 45.3 37.7 pH3.02 2.95 2.92 2.99 2.91 2.95 60° C. Doxorubicin·HCl assay 2.114 1.7521.393 1.176 0.925 mg/ml % stability 100.0 82.9 65.9 55.7 43.8 pH 3.022.96 2.98 2.96 2.97

[0147] Table 15—Doxorubicin.HCl 2 mg/ml pH 3.0 Solution in 5% Dextrose.

[0148] Arrhenius approach. Pseudo-first order rate constants,

[0149] Arrhenius equation, calculated t_(90%) PSEUDO-FIRST ORDER RATECONSTANTS, OBSERVED VALUES (K_(obs)) Temperature K_(obs) × 10³ (1/days)Correlation Coefficient 35° C. 4.190 0.990 45° C. 14.55 0.995 55° C.58.11 0.998 60° C. 102.6 0.999

[0150] Arrhenius Equation from 35° C., 45° C., 55° C. and 60° C. RateConstants

ln K _(obs)=−13266/T+37.56

[0151] correlation coefficient=0.9993 PSEUDO-FIRST ORDER RATE CONSTANTS,CALCULATED VALUES (K) Temperature K × 10³ (1/days) t_(90%) (days) 95%confidence limits  4° C. 0.0326 3,218 1,463-7,082  8° C. 0.0645 1,628  792-3,344 15° C. 0.203 516 281-949 27° C. 1.283 82  53-128

[0152] Long Term Stability of Doxorubicin Formulations having a pHFalling within the Range from 2.5 to 3.5

[0153] Batches tested, formulations tested and packaging used arereported on, respectively, tables 16, 17 and 18, as well as on,respectively, tables 25, 26 and 27.

[0154] Test and Methods

[0155] The formulations were tested as regards appearance, clarity ofsolution, pH, sterility (8° C., yearly), doxorubicin.HCl assay.

[0156] Test Methods

[0157] For appearance and clarity; visual inspection

[0158] For pH: USP XXI

[0159] For sterility; USP XXI (membrane filtration)

[0160] For doxorubicin.HCl assay: HPLC ion-pair method and USP HPLCmethod (USP XXI)

[0161] Brief description of the HPLC ion-pair method for doxorubicin.

[0162] HCl assay:

[0163] Column filling: reverse phase, Zortax TMS

[0164] Mobile phase: water, acetonitrile, methanol (54:29:17 v/v/v)containing 2 ml/l 85% phosphoric acid and 1 mg/ml sodium laurylsulfate(pairing agent) adjusted to pH 3.5 with 2N NaOH

[0165] Mobile phase flow rate: 1.5 ml/min

[0166] Column temperature: ambient (22° C.±2° C.)

[0167] Analytical wavelength: 254 nm

[0168] System suitability parameters: Symmetry factor between 0.7 and1.2; number of theoretical plates ≧2500; measurement reproducibility:variation coefficient <1, n=6; resolution factor ≧12

[0169] The HPLC ion-pair method for doxorubicin.HCl assay is validatedfor accuracy, precision, linearity, sensitivity, specificity andstability-indicating nature.

[0170] The results obtained for:

[0171] percent doxorubicin.HCl stability (ion-pair method) and

[0172] pH

[0173] referred to the vials stored in upright position are given in:

[0174] Table 19 storage at −20° C.

[0175] Tables 20 and 28 storage at +4° C.

[0176] Tables 21 and 29 storage at +8° C.

[0177] Tables 22 and 30 storage at +15° C.

[0178] Tables 23 and 31 storage at +27° C.

[0179] Table 24 storage at 100 and 250 foot candles

[0180] Table 32 storage at 250 foot candles.

[0181] The doxorubicin.HCl assays given in these tables are the mean ofthree independent determinations.

[0182] As far as the other parameters checked during stability:

[0183] the clarity of the solution was unchanged at all the checkscarried out at all the storage conditions applied;

[0184] the appearance of the solutions was: a) unchanged at all thechecks carried out on samples stored at 4° C. and 8° C., b) slightlydarkened after: 9 months at 15° C., 3 months at 27° C., 3 months at 100and 250 foot candles light;

[0185] the closure system was unchanged at all the checks carried out atall the storage conditions;

[0186] the sterility was maintained after 18 months at 8° C.

[0187] The results of the controls carried out on the vials stored ininverted position do not differ significantly from those on the vials inupright position.

[0188] The percent doxorubicin.HCl stability values obtained by the USPHPLC method do not differ significantly from those obtained by the HPLCion-pair method given in Tables 19-24.

[0189] The obtained stability data indicate that the testeddoxorubicin.HCl solutions having different pH values within the rangefrom 2.5 to 3.5 reach the lower limit of acceptance (90% of nominalconcentration) in about 9 and 2-3 months at 15° C. and, respectively 27°C., but prove stable up to 18 months at 4° C. and 8° C., i.e. attemperature usually adopted for the storage of the products underrefrigeration.

[0190] In distinct contrast, the doxorubicin.HCl solution obtained uponreconstitution of the commercial freeze-dried preparate, whose pH variesbetween 4.5 and 6, shows a much lower degree of stability as shown bythe fact that it is recommended to discard reconstituted solutions afteronly 48 hours storage in refrigerator according to the leafletaccompanying packages of Adriamycin (i.e. doxorubicin.HCl) in the UnitedStates. TABLE 16 Stability studies. Batches tested. Batch Batch NoCharacteristics TF/23049 TF/23077 TF/23078 TF/23117 TF/23119 H0001 L0001M0001 Doxorubicin·HCl 10 10 10 20 50 10 20 50 per vial (mg) pH 3.06 2.813.50 2.97 3.08 3.15 3.05 3.20 Formulation No. F16804/IL1 F16804/IL1F16804/IL1 F16804/IL2 F16804/IL3 F16804/IL4 F16804/IL5 F16804/IL6 Batchsize 700 400 400 500 500 2,400 2,300 2,400 No. of vials

[0191] TABLE 17 Stability studies. Formulations tested. Composition perFormulation number vial F16804/IL F16804/IL2 F16804/IL3 F16804/IL4F16804/IL5 F16804/IL6 Doxorubicin·HCl 10.0 20 50 10 20 50 mgHydrochloric acid 2.8-3.5 2.8-3.5 2.8-3.5 2.8-3.5 2.8-3.5 2.8-3.5 q.s.to pH Water q.s. to ml 5.0 10.0 25.0 5.0 10.0 25.0

[0192] TABLE 18 Stability studies. Packaging used. Batch No. PackagingTF/23049 TF/23077 TF/23078 TF/231117 TF/23119 H0001 L0001 M0001 vialglass type I I I I I I I I vial top 8 ml 8 ml 8 ml 14 ml 39 ml 10 ml 14ml 39 ml capacity stopper chlorobutyl chlorobutyl chlorobutylchlorobutyl chlorobutyl chlorobutyl chlorobutyl chlorobutyl rubber,rubber, rubber, rubber, rubber, rubber, rubber, rubber, teflon-facedteflon-faced teflon-faced teflon-faced teflon-faced teflon-facedteflon-faced teflon-faced seal aluminum aluminum aluminum aluminumaluminum aluminum aluminum aluminum

[0193] TABLE 19 Doxorubicin·HCl 2 mg/ml solution. Stability data at −20°C. (vials stored upright) acquired up to 3 months. Batch Time - MonthsDosage 0 1 3 H0001 doxorubicin·HCl 100 99.9 99.6 10 mg % stability pH3.15 3.12 2.98 L0001 doxorubicin·HCl 100 100.8 99.8 20 mg % stability pH3.05 2.84 2.97 M0001 doxorubicin·HCl 100 100.7 101.0 50 mg % stabilitypH 3.20 2.96 2.99

[0194] TABLE 20 Doxorubicin·HCl 2 mg/ml solution. Stability data at 4°C. (vials stored upright) acquired up to 18 months Batch Time MonthsDosage 0 1 3 6 9 12 18 TF/23049 * 100 99.9 100.6 98.3 98.2 97.7 96.9 10mg ** 3.06 3.10 3.09 3.10 3.05 2.97 3.07 TF/23077 * 100 101.7 99.3 97.998.0 99.8 10 mg ** 2.81 2.86 2.75 2.65 2.67 2.76 TF/23078 * 100 101.298.8 97.8 98.8 96.8 10 mg ** 3.50 3.54 3.49 3.44 3.43 3.54 TF/23117 *100 96.8 96.6 98.1 98.8 97.5 20 mg ** 2.97 2.98 2.92 2.86 2.95 2.98TF/23119 * 100 98.6 99.1 98.9 98.4 97.5 50 mg ** 3.08 2.98 2.98 2.892.99 3.00 H0001 * 100 97.6 99.2 10 mg ** 3.15 n.d. 3.06 3.22 L000 * 10098.8 98.4 20 mg ** 3.05 n.d. 2.99 2.94 M0001 * 100 99.7 99.7 50 mg **3.20 n.d. 3.00 3.04

[0195] TABLE 21 Doxorubicin·HCl 2 mg/ml solution. Stability data at 8°C. (vials stored upright) acquired up to 18 months Batch Time (Months)Dosage 0 1 2 3 6 9 12 18 TF/23049 * 100 99.7 100.1 96.5 96.1 96.5 95.410 mg ** 3.06 3.07 3.09 3.07 3.04 2.96 3.04 TF/23077 * 100 102.1 101.697.5 96.6 95.0 10 mg ** 2.81 2.81 2.74 2.65 2.67 2.75 TF/23078 * 10098.3 97.7 96.5 95.9 98.8 10 mg ** 3.50 3.59 3.47 2.27 3.43 3.51TF/23117 * 100 95.7 95.8 97.8 96.2 95.5 20 mg ** 2.97 2.97 2.92 2.852.96 2.98 TF/23119 * 100 97.6 97.8 96.2 97.3 96.8 50 mg ** 3.08 2.942.94 2.87 2.99 3.00 H0001 * 100 98.2 99.4 96.4 96.7 10 mg ** 3.15 3.123.16 3.05 3.23 L000 * 100 100.6 99.1 98.1 98.3 20 mg ** 3.05 2.84 2.832.97 2.94 M0001 * 100 100.3 100.6 98.7 99.0 50 mg ** 3.20 2.96 2.97 3.013.03

[0196] TABLE 22 Doxorubicin·HCl 2 mg/ml solution. Stability data at 15°C. (vials stored upright) acquired up to 12 months Batch Time (Months)Dosage 0 0.5 1 2 3 6 9 12 TF/23049 * 100 97.8 98.9 97.1 92.7 92.9 90.210 mg ** 3.06 3.03 3.07 3.10 3.08 3.02 2.95 TF/23077 * 100 100.4 101.998.8 94.6 92.7 91.1 10 mg ** 2.81 2.81 2.85 2.71 2.63 2.67 2.74TF/23078 * 100 101.4 98.4 95.3 94.6 91.9 90.7 10 mg ** 3.50 3.51 3.583.47 3.38 3.41 3.47 TF/23117 * 100 99.1 96.4 95.2 94.6 90.7 20 mg **2.97 2.95 2.95 2.90 2.81 2.95 TF/23119 * 100 97.4 97.1 95.9 92.7 90.6 50mg ** 3.08 2.99 2.95 2.91 2.87 2.98 H0001 * 100 97.9 97.1 94.8 94.6 10mg ** 3.15 3.12 3.16 3.06 3.23 L000 * 100 100.5 98.7 96.3 95.5 20 mg **3.05 2.85 2.87 2.98 2.96 M0001 * 100 99.4 100.3 97.2 95.6 50 mg ** 3.202.96 2.94 3.01 3.04

[0197] TABLE 23 Doxorubicin·HCl 2 mg/ml solution. Stability data at 27°C. (vials stored upright) acquired up to 6 months. Batch Time (Months)Dosage 0 0.5 1 2 3 6 TF/23049 * 100 97.2 95.8 87.9 73.6 10 mg ** 3.062.98 3.07 3.08 3.03 TF/23077 * 100 98.5 96.2 86.4 69.2 10 mg ** 2.812.80 2.85 2.71 2.64 TF/23078 * 100 101.2 94.5 80.5 71.1 10 mg ** 3.503.51 3.58 3.38 3.13 TF/23117 * 100 97.4 93.2 81.9 66.6 20 mg ** 2.972.95 2.94 2.88 2.77 TF/23119 * 100 96.0 93.3 85.3 66.8 50 mg ** 3.082.97 2.97 2.91 2.82 H0001 * 100 94.5 94.2 86.6 10 mg ** 3.15 3.10 3.093.01 L000 * 100 97.2 94.3 89.3 20 mg ** 3.05 2.84 2.85 2.96 M0001 * 10096.5 93.6 88.1 50 mg ** 3.20 2.95 2.95 2.99

[0198] TABLE 24 Doxorubicin·HCl 2 mg/ml solution. Stability data at 100and 250 f.c. (vials stored inverted) acquired up to 3 months. 100foot-candles 250 foot-candles Batch Time Months Dosage 0 0.5 1 3 0.5 1 23 TF/23049 * 100 96.3 95.9 81.3 95.9 94.8 10 mg ** 3.06 3.05 3.05 3.062.99 3.04 TF/23077 * 100 98.3 98.1 87.7 97.3 94.5 10 mg ** 2.81 2.792.84 2.70 2.79 2.84 TF/23078 * 100 99.6 96.4 88.0 97.8 89.7 10 mg **3.50 3.50 3.58 3.39 3.47 3.53 TF/23117 * 100 96.8 96.7 91.7 98.1 94.6 20mg ** 2.97 2.93 2.95 2.87 2.9 2.93 TF/23119 * 100 96.9 96.7 89.6 96.495.0 50 mg ** 3.08 2.96 2.95 2.93 2.96 2.97 H0001 * 100 95.2 93.7 87.810 mg ** 3.15 3.10 3.06 2.97 L000 * 100 96.5 93.0 86.5 20 mg ** 3.052.84 2.85 2.97 M0001 * 100 97.8 91.5 85.3 50 mg ** 3.20 2.95 2.94 2.99

[0199] TABLE 25 Stability studies. Batches tested. Batch Batch No.Characteristics P0001 Q0001 R0001 Doxorubicin·HCl 10 20 50 per vial (mg)pH 3.00 3.00 3.00 Formulation No. FI6804/IL7 FI6804/IL8 FI6804/IL9 Batchsize 2,400 2,200 2,500 No. of vials

[0200] TABLE 26 Stability studies. Formulations tested. Composition perFormulation number vial F16804/IL7 F16804/IL8 F16804/IL9 Doxorubicin·HCl10 20 50 per vial (mg) Hydrochloric acid 2.8-3.5 2.8-3.5 2.8-3.5 q.s. topH 0.9% sodium chloride 5.0 10.0 25.0 injection q.s. to ml

[0201] TABLE 27 Stability studies. Packaging used. Batch No PackagingP0001 Q0001 R0001 vial glass type I I I vial top 10 ml 14 ml 39 mlcapacity stopper chlorobutyl chlorobutyl chlorobutyl rubber, rubber,rubber, teflon-faced teflon-faced teflon-faced seal aluminum aluminumaluminum

[0202] TABLE 28 Doxorubicin·HCl 2 mg/ml solution in Saline for Injectionat pH = 3. Stability data at 4° C. (vials stored upright) acquired up to9 and 12 months. Batch Time - Months Dosage 0 3 6 9 12 P0001doxorubicin·HCl 100 98.3 98.0 99.2 10 mg % stability pH 3.00 2.93 2.982.90 Q0001 doxorubicin·HCl 100 97.5 97.0 100.1 20 mg % stability pH 3.013.06 3.03 3.00 R0001 doxorubicin·HCl 100 99.8 100.7 101.2 101.7 50 mg %stability pH 3.02 3.08 3.15 3.14 3.10

[0203] TABLE 29 Doxorubicin·HCl 2 mg/ml solution in Saline for Injectionat pH = 3. Stability data at 8° C. (vials stored upright) acquired up to9 and 12 months. Batch Time - Months Dosage 0 1 2 3 6 9 12 P0001doxorubicin·HCl 100 101.0 100.6 97.9 97.4 96.8 10 mg % stability pH 3.002.93 2.89 2.91 3.00 2.90 Q0001 doxorubicin·HCl 100 99.4 99.9 96.8 96.795.7 20 mg % stability pH 3.01 3.02 3.01 3.05 3.02 3.00 R0001doxorubicin·HCl 100 99.8 99.8 98.4 98.5 99.5 100.9 50 mg % stability pH3.02 3.02 3.09 3.08 3.13 3.13 3.10

[0204] TABLE 30 Doxorubicin·HCl 2 mg/ml solution in Saline for Injectionat pH = 3. Stability data at 15° C. (vials stored upright) acquired upto 9 and 12 months. Batch Time - Months Dosage 0 1 2 3 6 9 12 P0001doxorubicin·HCl 100 100.6 99.9 95.9 94.0 89.1 10 mg % stability pH 3.002.93 2.89 2.90 2.99 2.90 Q0001 doxorubicin·HCl 100 98.6 97.8 95.1 96.489.8 20 mg % stability pH 3.01 3.01 3.01 3.04 3.01 3.00 R0001doxorubicin·HCl 100 98.8 97.5 97.6 94.7 96.0 94.5 50 mg % stability pH3.02 3.02 3.08 3.08 3.14 3.11 3.10

[0205] TABLE 31 Doxorubicin·HCl 2 mg/ml solution in Saline for Injectionat pH = 3. Stability data at 27° C. (vials stored upright) acquired upto 3 months. Batch Time - Months Dosage 0 1 2 3 P0001 doxorubicin·HCl100 98.3 95.0 84.9 10 mg % stability pH 3.00 2.93 2.89 2.88 Q0001doxorubicin·HCl 100 98.0 93.2 83.8 20 mg % stability pH 3.01 3.01 2.993.03 R0001 doxorubicin·HCl 100 95.6 92.2 88.7 50 mg % stability pH 3.023.02 3.06 3.05

[0206] TABLE 32 Doxorubicin·HCl 2 mg/ml solution in Saline for Injectionat pH = 3. Stability data at R.T. + 250 f.c. (vials stored upright)acquired up to 3 months. Batch Time - Months Dosage 0 1 2 3 P0001doxorubicin·HCl 100 89.6 86.5 70.3 10 mg % stability pH 3.00 2.92 2.862.84 Q0001 doxorubicin·HCl 100 91.1 84.5 72.7 20 mg % stability pH 3.012.99 2.97 2.98 R0001 doxorubicin·HCl 100 96.0 91.4 86.6 50 mg %stability pH 3.02 3.01 3.04 3.02

[0207] TABLE 33 Stability Data of Doxorubicin Solution, 2 mg/ml and a pHof 3.0 at 45° C. Stabilizing Agent % Initial and Its Concentration 1 Wk2 Wk 4 Wk 8 Wk Water 87.8 75.9 53.8 25.5 5% Dextrose 91.1 82.3 65.6 38.85% Galactose 91.5 86.1 64.3 — 5% Fructose 91.9 80.6 64.1 — 4%α-L(−)-Fucose 91.2 81.9 63.8 — 4% α-D(+)-Fucose 91.8 81.9 63.3 — 1%Lactose 91.3 81.7 64.5 34.8 4% Dextran, MW 9,000 90.5 81.5 — — 4%Dextran, MW 506,000 92.0 84.0 — — 4% α-Cyclodextrin 91.7 84.3 — — 4%β-Cyclodextrin 92.1 84.1 — — 4% γ-Cyclodextrin 94.3 89.0 — — 5% Mannitol90.7 81.4 65.8 41.1 5% Sorbitol 91.4 83.0 67.2 42.5 0.5% Thioglycerol90.8 83.2 63.5 — 5% Inositol 91.7 84.9 — — 5% Ethanol 92.2 85.6 — — 10%Glycerol 92.2 83.4 65.5 —

[0208] The invention now being fully described, it will be apparent toone of ordinary skill in the art that many changes and modifications canbe made thereto without departing from the spirit or scope of theinvention as set forth herein.

What is claimed as new and desired to be secured by Letters of patent ofthe United States is: 1-17. (Cancelled)
 18. A physiologically acceptablesolution of doxorubicin hydrochloride dissolved in a physiologicallyacceptable solvent, having a pH adjusted to from about 2.5 to 5.0 with aphysiologically acceptable acid and has a concentration of doxorubicinof from 0.1 to 100 mg/ml.
 19. The physiologically acceptable solution ofdoxorubicin hydrochloride of claim 18, wherein said solution has notbeen reconstituted from a lyophilizate.
 20. The physiologicallyacceptable solution of doxorubicin hydrochloride of claim 18, whereinsaid solution is contained in a sealed container.
 21. Thephysiologically acceptable solution of doxorubicin hydrochloride ofclaim 18, wherein said solution has not been reconstituted from alyophilizate and is contained in a sealed container.
 22. Aphysiologically acceptable solution of doxorubicin hydrochloridedissolved in a physiologically acceptable solvent, having a pH adjustedto from 2.5 to 5.0 with a physiologically acceptable acid selected fromthe group consisting of hydrochloric acid, hydrobromic acid, sulfuricacid, phosphoric acid, nitric acid, acetic acid, succinic acid, tartaricacid, ascorbic acid, citric acid, glutamic acid, benzoic acid, methanesulfonic acid, and ethane sulfonic acid and the concentration of saiddoxorubicin hydrochloride being from 0.1 to 100 mg/ml.
 23. Thephysiologically acceptable solution of doxorubicin hydrochloride ofclaim 22, wherein said solution has not been reconstituted from alyophilizate.
 24. The physiologically acceptable solution of doxorubicinhydrochloride of claim 22, wherein said solution is contained in asealed container.
 25. The physiologically acceptable solution ofdoxorubicin hydrochloride of claim 22, wherein said solution has notbeen reconstituted from a lyophilizate and is contained in a sealedcontainer.
 26. The physiologically acceptable solution of doxorubicinhydrochloride of claim 18 or 22, wherein the physiologically acceptablesolvent is selected from the group consisting of water, physiologicalsaline, dextrose, polyethylene glycol, N,N-dimethylacetamide,N-hydroxy-2-ethyl-lactamide, ethanol, benzyl alcohol, propylene glycol,glycerin, diacetine, triacetine, polyethylene glycol 400, propyleneglycol methylether, isopropylidenglycerin, dimethylisosorbide,2-pyrrolidone, N-methyl-2-pyrrolidone, Brij®, Cremophor®, Myrj®, Tween®and Pluronics® and mixtures thereof.
 27. The physiologically acceptablesolution of doxorubicin hydrochloride of claim 18 or 22, wherein thephysiologically acceptable solvent is water.
 28. The physiologicallyacceptable solution of doxorubicin hydrochloride of claim 18 or 22,wherein said physiologically acceptable solvent is a saline solution.29. The physiologically acceptable solution of doxorubicin hydrochlorideof claim 18 or 22, wherein said physiologically acceptable solvent is adextrose solution.
 30. The physiologically acceptable solution ofdoxorubicin hydrochloride of claim 18 or 22, wherein saidphysiologically acceptable solvent is sterile water.
 31. Thephysiologically acceptable solution of doxorubicin hydrochloride ofclaim 18 or 22, further comprising a tonicity adjusting agent.
 32. Thephysiologically acceptable solution of doxorubicin hydrochloride ofclaim 18 or 22, wherein the concentration of doxorubicin hydrochlorideis from 0.1 to 50 mg/ml.
 33. The physiologically acceptable solution ofdoxorubicin hydrochloride of claim 18 or 22, wherein the concentrationof doxorubicin hydrochloride is from 1 to 20 mg/ml.
 34. Thephysiologically acceptable solution of doxorubicin hydrochloride ofclaim 18 or 22, wherein the pH of said solution is from about 2.5 toabout 3.5.
 35. The physiologically acceptable solution of doxorubicinhydrochloride of claim 18 or 22, wherein the pH of said solution is fromabout 2.7 to about 3.3.
 36. The physiologically acceptable solution ofdoxorubicin hydrochloride of claim 18 or 22, wherein said solution isstorage stable, intravenously injectable, sterile and pyrogen-free. 37.A physiologically acceptable solution of anthracycline glycosideselected from the group consisting of doxorubicin hydrochloride,epirubicin hydrochloride and idarubicin hydrochloride dissolved in aphysiologically acceptable solvent, having a pH adjusted to from about2.5 to 5.0 with a physiologically acceptable acid and has aconcentration of idarubicin of from 0.1 to 100 mg/ml.
 38. Thephysiologically acceptable solution of anthracycline glycoside of claim37, wherein said solution has not been reconstituted from alyophilizate.
 39. The physiologically acceptable solution ofanthracycline glycoside of claim 37, wherein said solution is containedin a sealed container.
 40. The physiologically acceptable solution ofanthracycline glycoside of claim 37, wherein said solution has not beenreconstituted from a lyophilizate and is contained in a sealedcontainer.
 41. A physiologically acceptable solution of anthracyclineglycoside selected from the group consisting of doxorubicinhydrochloride, epirubicin hydrochloride and idarubicin hydrochloridedissolved in a physiologically acceptable solvent, having a pH adjustedto from 2.5 to 5.0 with a physiologically acceptable acid selected fromthe group consisting of hydrochloric acid, hydrobromic acid, sulfuricacid, phosphoric acid, nitric acid, acetic acid, succinic acid, tartaricacid, ascorbic acid, citric acid, glutamic acid, benzoic acid, methanesulfonic acid, and ethane sulfonic acid and the concentration of saididarubicin hydrochloride being from 0.1 to 100 mg/ml.
 42. Thephysiologically acceptable solution of anthracycline glycoside of claim41, wherein said solution has not been reconstituted from alyophilizate.
 43. The physiologically acceptable solution ofanthracycline glycoside of claim 41, wherein said solution is containedin a sealed container.
 44. The physiologically acceptable solution ofanthracycline glycoside of claim 41, wherein said solution has not beenreconstituted from a lyophilizate and is contained in a sealedcontainer.
 45. The physiologically acceptable solution of anthracyclineglycoside of claim 37 or 41 wherein the physiologically acceptablesolvent is selected from the group consisting of water, physiologicalsaline, dextrose, polyethylene glycol, N,N-dimethylacetamide,N-hydroxy-2-ethyl-lactamide, ethanol, benzyl alcohol, propylene glycol,glycerin, diacetine, triacetine, polyethylene glycol 400, propyleneglycol methylether, isopropylidenglycerin, dimethylisosorbide,2-pyrrolidone, N-methyl-2-pyrrolidone, Brij®, Cremophor®, Myrj®, Tween®and Pluronics® and mixtures thereof.
 46. The physiologically acceptablesolution of anthracycline glycoside of claim 37 or 41 wherein thephysiologically acceptable solvent is water.
 47. The physiologicallyacceptable solution of anthracycline glycoside of claim 37 or 41 whereinsaid physiologically acceptable solvent is a saline solution.
 48. Thephysiologically acceptable solution of anthracycline glycoside of claim37 or 41 wherein said physiologically acceptable solvent is a dextrosesolution.
 49. The physiologically acceptable solution of anthracyclineglycoside of claim 37 or 41 wherein said physiologically acceptablesolvent is sterile water.
 50. The physiologically acceptable solution ofanthracycline glycoside of claim 37 or 41 further comprising a tonicityadjusting agent.
 51. The physiologically acceptable solution ofanthracycline glycoside of claim 37 or 41 wherein the concentration ofanthracycline glycoside selected from the group consisting ofdoxorubicin hydrochloride, epirubicin hydrochloride and idarubicinhydrochloride is from 0.1 to 50 mg/ml.
 52. The physiologicallyacceptable solution of anthracycline glycoside of claim 37 or 41 whereinthe concentration of anthracycline glycoside selected from the groupconsisting of doxorubicin hydrochloride, epirubicin hydrochloride andidarubicin hydrochloride is from 1 to 20 mg/ml.
 53. The physiologicallyacceptable solution of anthracycline glycoside of claim 37 or 41 whereinthe pH of said solution is from about 2.5 to about 3.5.
 54. Thephysiologically acceptable solution of anthracycline glycoside of claim37 or 41 wherein the pH of said solution is from about 2.7 to about 3.3.55. The physiologically acceptable solution of anthracycline glycosideof claim 37 or 41 wherein said solution is storage stable, intravenouslyinjectable, sterile and pyrogen-free.